Recovering What Matters : High Protein Recovery after Endotoxin Removal from LPS-Contaminated Formulations Using Novel Anti-Lipid A Antibody Microparticle Conjugates
Casonato Melo, Cristiane (Paris Lodron University of Salzburg. Department of Biosciences & Medical Biology)
Fux, Alexandra C. (Paris Lodron University of Salzburg. Department of Biosciences & Medical Biology)
Himly, Martin (Paris Lodron University of Salzburg. Department of Biosciences & Medical Biology)
Bastús, Neus G (Institut Català de Nanociència i Nanotecnologia)
Schlahsa, Laura (Miltenyi Biotec B.V. & Co)
Siewert, Christiane (Miltenyi Biotec B.V. & Co)
Puntes, Víctor (Institut Català de Nanociència i Nanotecnologia)
Duschl, Albert (Paris Lodron University of Salzburg. Department of Biosciences & Medical Biology)
Gessner, Isabel (Miltenyi Biotec B.V. & Co)
Fauerbach, Jonathan A. (Miltenyi Biotec B.V. & Co)

Data:	2023
Resum:	Endotoxins or lipopolysaccharides (LPS), found in the outer membrane of Gram-negative bacterial cell walls, can stimulate the human innate immune system, leading to life-threatening symptoms. Therefore, regulatory limits for endotoxin content apply to injectable pharmaceuticals, and excess LPS must be removed before commercialization. The majority of available endotoxin removal systems are based on the non-specific adsorption of LPS to charged and/or hydrophobic surfaces. Albeit effective to remove endotoxins, the lack of specificity can result in the unwanted loss of essential proteins from the pharmaceutical formulation. In this work, we developed microparticles conjugated to anti-Lipid A antibodies for selective endotoxin removal. Anti-Lipid A particles were characterized using flow cytometry and microscopy techniques. These particles exhibited a depletion capacity > 6 ×10 3 endotoxin units/mg particles from water, as determined with two independent methods (Limulus Amebocyte Lysate test and nanoparticle tracking analysis). Additionally, we compared these particles with a non-specific endotoxin removal system in a series of formulations of increasing complexity: bovine serum albumin in water < insulin in buffer < birch pollen extracts. We demonstrated that the specific anti-Lipid A particles show a higher protein recovery without compromising their endotoxin removal capacity. Consequently, we believe that the specificity layer integrated by the anti-Lipid A antibody could be advantageous to enhance product yield.
Ajuts:	European Commission 812661
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	LPS contamination ; Polystyrene particles ; Bioconjugation ; Supramolecular structures ; LAL ; NTA
Publicat a:	International journal of molecular sciences, Vol. 24, Issue 18 (September 2023) , art. 13971, ISSN 1422-0067

DOI: 10.3390/ijms241813971
PMID: 37762274

15 p, 2.2 MB

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